公开(公告)号：US06368830B1

公开(公告)日：2002-04-09

申请号：US09671950

申请日：2000-09-27

申请人： David J. Lampe ,  Hugh M. Robertson ,  Eric J. Rubin ,  Brian J. Akerley

发明人： David J. Lampe ,  Hugh M. Robertson ,  Eric J. Rubin ,  Brian J. Akerley

IPC分类号： C12N1574

CPC分类号： C12N9/22 ,  C12N15/74

摘要： Mariner-family transposable elements are active in a wide variety of organisms and are becoming increasingly important genetic tools in species lacking sophisticated genetics. The Himar1 element, a member of the mariner family, isolated from the horn fly, Haematobia irritans, is active in Escherichia coli when expressed appropriately. Using this fact, a genetic screen was devised to isolate hyperactive mutants of Himar1 transposase that enhance overall transposition from 4 to 50-fold as measured in an E. coli assay. These hyperactive Himar1 mutant transposases should enable sophisticated analysis of the biochemistry of mariner transposition and should improve efficiency of a variety of genetic manipulations involving transposition in vivo and in vitro such as random mutagenesis or transgenesis in a wide range of host cells than the transposable elements previously available.

摘要翻译： 水手家族转座因子在各种生物体中是活跃的，并且在缺乏复杂遗传学的物种中变得越来越重要的遗传工具。 Himar1元素，海员家族的成员，从角飞，哈马氏杆菌分离，在适当表达时在大肠杆菌中活跃。 使用这个事实，设计了一种遗传筛选来分离Himar1转座酶的活性突变体，其将整体转座增加4倍至50倍，如在大肠杆菌测定中所测定的。 这些超活跃的Himar1突变体转座酶应该能够复杂地分析海员转座的生物化学，并且应该提高涉及体内和体外转座的各种遗传操作的效率，例如在广泛的宿主细胞中的随机诱变或转基因，而不是以前的转座元件 可用。